Recombinant expression of Garlic virus C (GARV-C) capsid protein in insect cells and its potential for the production of specific antibodies

被引:7
作者
Alves-Junior, Miguel [1 ,2 ]
Marraccini, Fernanda Menezes [1 ]
Melo Filho, Pericles de Albuquerque [2 ]
Dusi, Andre Nepomuceno [3 ]
Pio-Ribeiro, Gilvan [2 ]
Ribeiro, Bergmann Morais [1 ]
机构
[1] Univ Brasilia, Dept Cell Biol, Brasilia, DF, Brazil
[2] Univ Fed Rural Pernambuco, Dept Agron, Recife, PE, Brazil
[3] Embrapa Hortalicas, Gama, DF, Brazil
关键词
capsid protein; GARV-C; allium spp; Allexivirus;
D O I
10.1016/j.micres.2006.06.016
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Garlic cultivars in Brazil are infected by a complex of viruses and for some virus species, such as the allexivirus, purification of the virions is sometimes cumbersume. To overcome this problem, recombinant expression of viral. proteins in heterologous systems is an alternative method for producing antibodies. The capsid gene from Garlic virus C (GarV-C), an Allexivirus, was inserted into the genome of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) generating the recombinant virus vSynGarV-C. The recombinant protein expression was confirmed by SDS-PAGE and western-blot of extracts from recombinant virus infected insect cells, where a protein band of approximately 32 KDa was observed only in extracts from recombinant infected cells. This protein corresponded to the predicted size of the capsid protein of the GarV-C. A rabbit polyclonal antibody was raised against this protein, shown to be specific for the GarV-C protein in western-blot and dot-Elisa, however with a low titer. (c) 2006 Elsevier GmbH. All rights reserved.
引用
收藏
页码:354 / 361
页数:8
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